Extracellular Vesicles Secreted by Human Urine-Derived Stem Cells Promote Ischemia Repair in a Mouse Model of HindLimb Ischemia
Extracellular Vesicles Secreted by Human Urine-Derived Stem Cells Promote Ischemia Repair in a Mouse Model of HindLimb Ischemia
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人尿干细胞分泌的细胞外囊泡促进小鼠后肢缺血模型的缺血修复
DOI：
10.1159/000490214
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发表时间：
2018
影响因子：
--
通讯作者：
Deng Zhifeng
中科院分区：
文献类型：
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作者：
Zhu Qingwei;Li Qing;Niu Xin;Zhang Guowei;Ling Xiaozheng;Zhang Jieyuan;Wang Yang;Deng Zhifeng
背景/目的我们的前期研究表明，人尿源性干细胞（USCs）作为细胞治疗和组织工程的细胞来源具有巨大的潜力，并且USCs分泌的细胞外囊泡（USCs-EVs）在动物模型中可以预防糖尿病诱导的肾损伤。本研究旨在评估USCs-EVs对缺血修复的影响。MethodsUSCs-EVs通过离心和过滤步骤的电池分离和纯化。然后通过透射电子显微镜、蛋白质印迹和可调电阻脉冲传感技术对USCs-EV进行表征。将USCs-EV肌内移植到缺血小鼠后肢后，我们观察了USCs-EV对激光多普勒灌注成像的灌注，组织学和免疫组织化学技术的血管生成和肌肉再生的治疗作用超过21天。随后，我们通过细胞计数试剂盒8体外试验测试了USCs-EV是否可以诱导人微血管内皮细胞系HMEC-1和小鼠成肌细胞系C2 C12的增殖。结果USCs-EVs为球形囊泡，直径30-150 nm，表达CD 9、CD 63和Tsg 101等外泌体标志物。USCs-EV给药后，后肢缺血（HLI）模型中的缺血肢体灌注和功能显著增加。此外，USCs-EV治疗组中的血管生成和肌肉再生水平显著高于PBS组。在体外实验表明，USCs-EVs促进HMEC-1和C2 C12细胞增殖的剂量依赖martens.ConclusionsThese结果首次揭示，USCs-EVs有效地减轻严重的后肢缺血性损伤，并代表一种新的治疗HLI。
Background/AimsOur previous studies have shown that human urine-derived stem cells (USCs) have great potential as a cell source for cytotherapy and tissue engineering and that extracellular vesicles (EVs) secreted by USCs (USCs-EVs) can prevent diabetes-induced kidney injury in an animal model. The present study was designed to evaluate the effects of USCs-EVs on ischemia repair.MethodsUSCs-EVs were isolated and purified by a battery of centrifugation and filtration steps. The USCs-EVs were then characterized by transmission electron microscopy, western blot and tunable resistive pulse sensing techniques. After intramuscularly transplanting USCs-EVs into an ischemic mouse hind-limb, we observed the therapeutic effects of USCs-EVs on perfusion by laser doppler perfusion imaging, angiogenesis and muscle regeneration by histology and immunohistochemistry techniques over 21 days. We subsequently tested whether USCs-EVs can induce the proliferation of a human microvascular endothelial cell line HMEC-1 and a mouse myoblast cell line C2C12 by cell counting kit 8 assay in vitro. Meanwhile, the potential growth factors in the USCs-EVs and supernatants of the USCs cultures were detected by enzyme-linked immunosorbent assay.ResultsThe USCs-EVs were spherical vesicles with a diameter of 30–150 nm and expressed exosomal markers, such as CD9, CD63 and Tsg101. Ischemic limb perfusion and function were markedly increased in the hind-limb ischemia (HLI) model after USCs-EVs administration. Moreover, angiogenesis and muscle regeneration levels were significantly higher in the USCs-EVs treatment group than in the PBS group. The in vitro experiments showed that USCs-EVs facilitated HMEC-1 and C2C12 cell proliferation in a dose-dependent manner.ConclusionsThese results revealed for the first time that USCs-EVs efficiently attenuate severe hind-limb ischemic injury and represent a novel therapy for HLI.